AU2011291626A1 - Synergistic algicidal compositions including hydrazone derivatives and copper - Google Patents

Abstract

The present invention relates to the use of mixtures containing hydrazone compounds and copper for controlling the growth of algae.

Description

WO 2012/024494 PCT/US2011/048260 SYNERGISTIC ALGICIDAL COMPOSITIONS INCLUDING HYDRAZONE DERIVATIVES AND COPPER Cross Reference to Related Applications [0001] This application claims the benefit of U.S. Provisional Patent Application Serial No. 61/375,471 filed August 20, 2010, which is expressly incorporated by reference herein. Field of the Invention [0002] The present invention relates to the use of hydrazones in combination with copper, copper-based algicides or other copper-containing materials as synergistic algicidal mixtures. Background [0003] The present invention relates to the use of hydrazone derivatives in combination with copper for controlling algae. In particular, the present invention relates to a method for controlling algae by the use of certain hydrazone derivatives when applied in synergistic mixtures with copper. Such mixtures of hydrazone derivatives with copper are known to exert a synergistic effect towards fungi. Examples include those disclosed in PCT Patent Application PCT/US10/021040, filed on t4 January 2010. [0004] The presence of algae and other fouling organisms in various aqueous systems or systems exposed to water such as lattices, paints, coatings, cooling water systems, the marine environment, and decorative ponds can cause deterioration or disfigurement of the system. For example, painted surfaces may be disfigured by the unsightly buildup of algae, detracting from the overall aesthetics of the painted article and cooling towers or boats may lose efficiency due to the buildup of algae on surfaces. It is conventional to practice methods which inhibit the algal deterioration of such systems by incorporating a variety of additives or combinations of additives that are characterized by having antialgal activity. [0005] A wide variety of materials have been used to control algae in different environments, including; chlorine/bromine compounds, glutaraldehyde, isothiazoles, organotin formulations, copper salts, quaternary ammonium compounds and triazines. However, each of these materials has deficiencies related to toxicity, pH and temperature sensitivity, limited effectiveness, chemical stability, and/or compatibility. Due to these 1 WO 2012/024494 PCT/US2011/048260 deficiencies in conventional antialgal compounds, there is a continuing need for more effective antialgal agents. [0006] Copper and copper products are widely used as aquatic biocides and antifouling agents in fresh or marine environments to control unwanted organisms based on the toxicity of copper towards algae, fungi, macrophytes and mollusks (Handbook of Copper Compounds and Applications, edited by H. W. Richardson and published by Marcel Dekker, Inc. New York (1997), which is expressly incorporated by reference herein). [0007] Ecological risk assessment studies have shown that copper products, which normally are applied at high use rates, may be toxic to birds, mammals, fish and other aquatic species ("Reregistration Eligibility Decision (RED) for Coppers," EPA 738-R-06-020, July 2006, which is expressly incorporated by reference herein). Thus, while copper is a highly useful agent for controlling the growth of undesirable organisms in different environments, it is desirable to minimize the amount of copper applied. [0008] We have discovered that hydrazone derivatives in combination with copper inhibit the growth of algae. Due to strong synergism between hydrazones and copper, the amount of copper traditionally needed to inhibit growth of algae can be greatly reduced in the presence of hydrazone derivatives. The present invention is a method for controlling algae, comprising applying to the locus of the algae an algicidally effective amount of one or more compounds of the structural formula I in combination with copper. Summary of the Invention [0009] One exemplary embodiment of the present disclosure includes a synergistic mixture for controlling the growth of fungi, the synergistic mixture including copper and a hydrazone compound of Formula I:

X

3 Y 3 X4 OH O24 SN,N W 5 H n

X

6 R Y6 Formula I wherein W is -CHR 1-; n is 0 or 1; 2 WO 2012/024494 PCT/US2011/048260 R is H, C 1

-C

6 alkyl, C 2

-C

6 alkenyl, C 2

-C

6 alkynyl, C 3

-C

6 cycloalkyl, CI-C 6 haloalkyl,

C

2

-C

6 haloalkenyl, C 2

-C

6 haloalkynyl, or C 3

-C

6 halocycloalkyl; RI is H, C 1

-C

6 alkyl, C 2

-C

6 alkenyl, C 2

-C

6 alkynyl, C 3

-C

6 cycloalkyl, CI-C 6 haloalkyl,

C

2

-C

6 haloalkenyl C 2

-C

6 haloalkynyl, C 3

-C

6 halocycloalkyl, substituted aryl, unsubstituted aryl, substituted heteroaryl, or unsubstituted heteroaryl, substituted benzyl, or unsubstituted benzyl;

X

3 , X 4 , X 5 , and X 6 are each independently selected from the group consisting of H, halogen, nitro, hydroxyl, cyano, CI-C 4 alkyl, CI-C 4 alkoxy, C 2

-C

4 alkenyl, C 2

-C

4 alkynyl, C 1 C 4 alkylthio, C 1

-C

4 haloalkyl, C 1

-C

4 haloalkoxy, C 2

-C

4 haloalkenyl, C 2

-C

4 haloalkynyl, C 1

-C

4 haloalkylthio, -SO 2 R1, -SONRIRI, -CR1=NOR1, -CONRIRI, -NR1COOR1, -COORI, -NR1R1, -NR1SO 2 R1, substituted aryl, substituted heteroaryl, substituted benzyl, substituted benzoyl, substituted phenoxy, unsubstituted aryl, and unsubstituted heteroaryl, unsubstituted benzyl, unsubstituted benzoyl, and unsubstituted phenoxy; and

Y

2 , Y 3 , Y 4 , Y 5 , and Y 6 are each independently selected from the group consisting of H, halogen, nitro, hydroxyl, cyano, C 1

-C

4 alkyl, C 1

-C

4 alkoxy, C 2

-C

4 alkenyl, C 2

-C

4 alkynyl, C 1 C 4 alkylthio, C 1

-C

4 haloalkyl, C 1

-C

4 haloalkoxy, C 2

-C

4 haloalkenyl, C 2

-C

4 haloalkynyl, C 1

-C

4 haloalkylthio, -SO 2 R1, -SONRIRI, -CR1=NOR1, -CONRIRI, -NR1COOR1, -COORI, -NR1R1, -NR1SO 2 R1, substituted aryl, substituted heteroaryl, substituted benzyl, substituted benzoyl, substituted phenoxy, unsubstituted aryl, unsubstituted heteroaryl, unsubstituted benzyl, unsubstituted benzoyl and unsubstituted phenoxy; with the proviso that X 3 and X 4 , X 4 and X 5 , X 5 and X 6 , Y 2 and Y 3 , or Y 3 and Y 4 may form a 5- or 6-membered fused ring which may contain up to two heteroatoms selected from the group consisting of 0, N, and S. [0010] The term "alkyl" refers to a branched, unbranched, or cyclic carbon chain, including methyl, ethyl, propyl, butyl, isopropyl, isobutyl, tertiary butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. [0011] The term "cycloalkyl" refers to a monocyclic or polycyclic, saturated substituent consisting of carbon and hydrogen. 3 WO 2012/024494 PCT/US2011/048260 [0012] The term "alkenyl" refers to a branched, unbranched or cyclic carbon chain containing one or more double bonds including ethenyl, propenyl, butenyl, isopropenyl, isobutenyl, cyclohexenyl, and the like. [0013] The term "alkynyl" refers to a branched or unbranched carbon chain containing one or more triple bonds including propynyl, butynyl and the like. [0014] As used throughout this specification, the term 'R' refers to the group consisting of C 1

-C

6 alkyl, C 2

-C

6 alkenyl, C 2

-C

6 alkynyl, C 3

-C

6 cycloalkyl, CI-C 6 haloalkyl,

C

2

-C

6 haloalkenyl, C 2

-C

6 haloalkynyl, or C 3

-C

6 halocycloalkyl, unless stated otherwise. [0015] The term "alkoxy" refers to an -OR substituent. [0016] The term "alkylthio" refers to an -SR substituent. [0017] The term "haloalkylthio" refers to an alkylthio, which is substituted with Cl, F, I, or Br or any combination thereof. [0018] The term "cyano" refers to a -C--N substituent. [0019] The term "hydroxyl" refers to an -OH substituent. [0020] The term "haloalkoxy" refers to an alkoxy substituent, which is substituted with Cl, F, Br, or I, or any combination thereof. [0021] The term "haloalkyl" refers to an alkyl, which is substituted with Cl, F, I, or Br or any combination thereof. [0022] The term "halocycloalkyl" refers to a monocyclic or polycyclic, saturated substituent consisting of carbon and hydrogen, which is substituted with Cl, F, I, or Br or any combination thereof. [0023] The term "haloalkenyl" refers to an alkenyl, which is substituted with Cl, F, I, or Br or any combination thereof. [0024] The term "haloalkynyl" refers to an alkynyl which is substituted with Cl, F, I, or Br or any combination thereof. [0025] The term "halogen" or "halo" refers to one or more halogen atoms, defined as F, Cl, Br, and I. [0026] The term "aryl" refers to a cyclic, aromatic substituent consisting of hydrogen and carbon. [0027] The term "heteroaryl" refers to a cyclic substituent that may be fully unsaturated, where the cyclic structure contains at least one carbon and at least one heteroatom, where said heteroatom is nitrogen, sulfur, or oxygen. [0028] The term "phenoxy" refers to an -O substituted with a six-membered fully unsaturated ring consisting of hydrogen and carbon. 4 WO 2012/024494 PCT/US2011/048260 [0029] The term "benzyl" refers to a -CH 3 substituted with a six-membered fully unsaturated ring consisting of hydrogen and carbon. [0030] The term "benzoyl" refers to a carbonyl substituted with a six-membered fully unsaturated ring consisting of hydrogen and carbon [0031] The term "nitro" refers to a -NO 2 substituent. [0032] Certain compounds disclosed in this document can exist as one or more isomers. It will be appreciated by those skilled in the art that one isomer may be more active than the others. The structures disclosed in the present disclosure are drawn in only one geometric form for clarity, but are intended to represent all geometric or tautomeric forms of the molecule. [0033] Additional features and advantages of the present invention will become apparent to those skilled in the art upon consideration of the following detailed description of the illustrative embodiments exemplifying the best mode of carrying out the invention as presently perceived. Detailed Description of the Disclosure [0034] The embodiments of the invention described herein are not intended to be exhaustive or to limit the invention to the precise forms disclosed. Rather, the embodiments selected for description have been chosen to enable one skilled in the art to practice the invention. Although the disclosure is described as a synergistic combination of copper, copper based algicides, or other copper-containing materials and a hydrazone or hydrazone derivative it should be understood that the concepts presented herein may be used in various applications and should not be limited. [0035] The mixtures of the present invention may have broad spectrum algicidal activity. Algae which may be controlled by the method of the present invention include individual species and mixed cultures. Examples of species controlled include green algae such as Chlamydomonas reinhardtii, Chlorella pyrenoidosa, Scenedesmus quadricauda, Chlorococcum oleofaciens, and Selenastrum species; blue-green algae (cyanobacteria) such as Phormidium species, Anabaenaflosaquae, Nostoc commune, Osiffiatorae species, Synechocystis species, and Synechococcus species; and marine algae such as Dunaliella parva. [0036] According to the method of the present invention, the mixtures described herein may be combined with other known antialgal compounds including: chlorine/bromine compounds; glutaraldehyde, isothiazoles, isothiazolones, organotin formulations, quaternary ammonium compounds, and triazines. 5 WO 2012/024494 PCT/US2011/048260 [0037] The amount of the active mixture required to control algae will depend upon many factors such as, for example: the type of surface; the amount of water present; whether the active mixture is incorporated into a coating composition, applied directly to an object, or added to an aqueous or other solution; and the type and extent of algal infestation. [0038] While the mixtures described herein may be administered alone to control algae, it is preferable to administer them as formulations. Useful formulations comprise one or more compounds and one or more acceptable carriers. The term "acceptable carrier" means a carrier compatible with the active mixture and other ingredients of the formulation and which is not toxic to the system or which will not cause degradation of the system. Formulations of the mixtures may contain from 0.01 to 99.9 percent by weight of the mixture. More typically the solutions and formulations will contain from 1.0 to 85 percent by weight of the mixture. Useful formulations include aqueous solutions, solvent based solutions, wettable powders, emulsifiable concentrates, dusts, granular formulations, pellets, aerosols, or flowable emulsion concentrates. In such formulations, the compounds are extended with liquid or solid carriers and, when desired, suitable surfactants are incorporated. [0039] In the case of spray formulations, it is often desirable to include one or more adjuvants, such as wetting agents, spreading agents, dispersing agents, stickers, adhesives, emulsifying agents and the like. Such adjuvants commonly used in the art can be found in the John W. McCutcheon, Inc. publication Detergents and Emulsifiers, Annual, Allured Publishing Company, Ridgewood, New Jersey, U.S.A. Spray formulations can be administered using common application methods, such as conventional high-volume hydraulic sprays, low volume sprays, air-blast spray, aerial sprays, backpack and hand held sprays, and dusts. The dilution and rate of application will depend upon the type of equipment employed, the method of application, area treated, and algae to be controlled. [0040] The mixtures of the present invention may also be used to control algae in cooling tower waters. In such applications the hydrazone and copper components of the mixtures are maintained at a concentration of from 0.001 ppm to the solubility limit of the compound, preferably 0.01 to 200 ppm. [0041] In addition, the compounds of the present invention are useful for imparting algal resistance to coatings or impregnated compositions. In such applications, the mixtures are incorporated into the coating or into the impregnating composition at a concentration from 0.1 to 10 percent by weight, preferably 1 to 5 percent by weight. [0042] The compounds of the present invention may also be useful for imparting algal resistance to construction products such as stucco, roof mastics, wall mastics, and 6 WO 2012/024494 PCT/US2011/048260 masonry coatings; in clear finishes and coatings to protect underlying substrates from algae; for algae control in aquaculture including aquaria, fish hatcheries, shrimp ponds, finfish ponds, mollusk and crustacean cultivation; for algae control in recreational and decorative bodies of water such as swimming pools, lakes, fountains, and decorative ponds; for algae control in bodies of water for industrial or municipal use, such as settling or separation ponds, waste treatment ponds, and water reservoirs; for algae control in hydroponic farming or rice paddies; for algae control in processing and manufacture of pulp and paper products; and for inclusion in plastics or in coatings for plastics to protect against algae. Care must be taken in the selection of compound and application rate to avoid adverse effects on non-target organisms. [0043] The mixtures have broad ranges of efficacy as algicides. The exact amounts of hydrazones and copper-containing materials to be applied is dependent not only on the specific materials being applied and relative amounts of hydrazone and copper in the mixtures, but also on the particular action desired, the algal species to be controlled, and the stage of growth thereof, as well as the location to be contacted with the mixture. Thus, all the mixtures, and formulations containing the same, may not be equally effective at similar concentrations or against the same algal species. [0044] The exact amount of a mixture required varies with the algal species to be controlled, the type of formulation employed, the method of application, climate conditions, and the like. The dilution and rate of application will depend upon the type of equipment employed, the method and frequency of application desired and algal species to be controlled. The molar ratio of copper to hydrazone in the active mixture may range from 0.1:1 to 10,000:1, preferably from 0.5:1 to 1000:1 and more preferably from 1:1 to 20:1. [0045] It should be understood that the preferred amount of a copper material to be mixed with hydrazone in a given application may be influenced by availability of copper from other sources such as copper present in the body of water or material to be treated, copper present in the water or solvent used in preparing the algicidal solutions for application such as in spray application, copper present in formulations used in preparing spray solutions or dusts for application, or any other suitable copper source. [0046] For algal control the hydrazone may be applied before or after the application of copper such that the mixture is generated in the location where control is desired. Additionally, multiple applications of copper or the hydrazone may be applied. [0047] As exemplified below, hydrazones of the present invention, or their metal complexes, in a mixture with inorganic or organic mono-or divalent copper salts or chelates 7 WO 2012/024494 PCT/US2011/048260 (hereinafter referred to as "copper products") increase the biological potency of copper products, enabling comparable or improved efficacy at lower copper use rates. While not intending to be all-inclusive, copper products which may be mixed with the compounds of the present invention to provide enhanced potency may include the following: copper oxychloride, copper octanoate, copper ammonium carbonate, copper arsenate, copper oxysulfate, copper formate, copper propionate, copper oxyacetate, copper citrate, copper chloride, copper diammonium chloride, copper nitrate, copper carbonate, copper phosphate, copper pyrophosphate, copper disodium EDTA, copper diammonium EDTA, copper oxalate, copper tartrate, copper gluconate, copper glycinate, copper glutamate, copper aspartate, copper adipate, copper palmitate, copper stearate, copper caprylate, copper decanoate, copper undecylenate, copper neodecanoate, copper linoleate, copper oleate, copper borate, copper methanesulfonate, copper sulfamate, copper acetate, copper hydroxide, copper oxide, copper oxychloride-sulfate, copper sulfate, basic copper sulfate, copper-oxine, copper 3 phenylsalicylate, copper chloride hydroxide, copper dimethyldithiocarbamate, ammonium copper sulfate, copper magnesium sulfate, copper naphthenate, copper ethanolamine, chromated copper arsenate, ammoniacal copper arsenate, ammoniacal copper zinc arsenate, ammoniacal copper borate, Bordeaux mixture, copper zinc chromate, cufraneb, cupric hydrazinium sulfate, cuprobam, nano-copper materials, copper didecyldimethylammonium chloride, Algimycin P11-C, Aquatrine, A&V-70, Cutrine-plus, Stocktrine II and K-Tea algicide and where appropriate the hydrates of such compounds. [0048] Methods for preparation of salicylaldehyde benzoylhydrazones and 2 hydroxyphenylketone benzoylhydrazones from salicylaldehydes or 2-hydroxyphenyl ketones and a benzoic acid hydrazide are well known in the literature. In addition the preparation of metal complexes of these materials is also well known (see for example Ainscough et al. J. Inorg. Biochem. 1999, 77, 125-133, which is expressly incorporated by reference herein). Methods of preparation of precursor hydrazides are also well known. Hydrazides can be prepared, for example, from carboxylic acids such as in Maxwell et al., J. Med. Chem. 1984, 27, 1565-1570, and from carboxylic esters such as in Dydio et al., J. Org. Chem. 2009, 74, 1525-1530, which are expressly incorporated by reference herein. Thus, the synthesis of any benzoylhydrazone of the present invention and its metal complex(es) is fully described where the starting aldehyde or ketone, and the starting benzoic acid hydrazide, benzoic acid, or benzoic acid ester are described. The hydrazones disclosed may also be in the form of algicidally acceptable salts and hydrates. Examples 8 and 9 below provide typical methods 8 WO 2012/024494 PCT/US2011/048260 for the preparation of such benzoylhydrazones. Example 12 below provides a general method for the preparation of their metal complexes [0049] The compounds of Formula I may, therefore, be made using well-known chemical procedures. Intermediates not specifically mentioned in this disclosure are either commercially available, may be made by routes disclosed in the chemical literature, or may be readily synthesized from commercial starting materials utilizing standard procedures. [0050] The following examples are presented to illustrate the various aspects of the compounds of the present disclosure and should not be construed as limitations to the claims. [0051] Example 1. Preparation of 2-hydroxy-3,5-bis trifluoromethylbenzaldehyde

CF

3 OH

CF

3 CHO 3,5-Bis(trifluoromethyl)anisaldehyde (prepared as in Sui and Macielag, Synth. Commun. 1997, 27, 3581-3590, which is expressly incorporated by reference herein; 2.0 grams (g), 7.7 millimoles (mmol)) was dissolved in dry dichloromethane (CH 2 Cl 2 ; 15 milliliters (mL)), cooled to -78 'C and treated in portions with boron tribromide (BBr 3 , 1 M solution in

CH

2 Cl 2 ; 8.0 mL, 8.0 mmol). The mixture was stirred and allowed to warm to 25 'C. After 20 hours (h), the mixture was cooled to -40 'C, carefully treated with H 2 0 (10 mL) and warmed to room temperature. The separated organic phase was washed with water (H 2 0; 10 mL), saturated (satd) sodium chloride (NaCl) solution (5 mL), dried over sodium sulfate (Na 2

SO

4 ) and evaporated. The residue was purified by silica gel chromatography with a 0 to 20% gradient of ethyl acetate (EtOAc) in hexane to give the purified aldehyde (1.4 g, 70%) as an oil: 1 H NMR (400 MHz, CDCl 3 ) 6 12.05 (s, 1H), 10.02 (s, 1H), 8.07 (s, 2 H). EIMS m/z 258. [0052] Example 2. Preparation of 5-chloro-2-hydroxy-3-trifluoromethyl benzaldehyde F F F OH CI CHO 5-Chloro-2-fluorobenzotrifluoride (1.5 g, 7.5 mmol) was dissolved in dry tetrahydrofuran (THF; 10 mL), treated with tetramethylethylenediamine (TMEDA; 1.6 mL, 1.2 g, 11 mmol), cooled to -78 'C and treated in portions with n-butyl lithium (n-BuLi, 2.5 M in hexanes; 3.9 mL, 9.8 mmol). After stirring at -78 'C for 90 minutes (min), the mixture was treated with 9 WO 2012/024494 PCT/US2011/048260 N,N-dimethylformamide (DMF; 770 microliters (pL), 730 mg, 10 mmol) and stirred for a further 30 min. The cooling bath was removed and mixture warmed to 25 'C over 30 min. The reaction was quenched by addition of satd ammonium chloride (NH 4 Cl) solution then diluted with diethyl ether (Et 2 0; 30 mL). The separated organic phase was washed with satd NaCl (10 mL), dried (Na 2

SO

4 ) and evaporated. The residue was dissolved in dry methanol

(CH

3 0H; 10 mL) and treated with 30% sodium methoxide solution in CH 3 0H (14 g). The mixture was stirred at 25 'C for 20 h, diluted with H 2 0 (50 mL) and extracted with Et 2 0 (2 x 40 mL). The combined organic phases were washed with satd NaCl solution (20 mL), dried (Na 2

SO

4 ) and evaporated. The residue was purified by silica gel chromatography using a 0 to 10% EtOAc gradient in hexane to give the benzaldehyde (1.1 g). This material (1.0 g, 4.2 mmol) was dissolved in dry CH 2 Cl 2 (10 mL), cooled to -78 'C and treated with BBr 3 (1 M solution in CH 2 Cl 2 ; 5 mL, 5 mmol). The mixture was allowed to warm to 25 'C and stirred for 22 h. After cooling to -45 C, the mixture was treated with H 2 0 (5 mL), warmed to 25 'C and extracted with EtOAc (2 x 15 mL). The combined extracts were washed with satd NaCl solution (10 mL), dried (Na 2

SO

4 ) and evaporated. The residue was purified by silica gel chromatography using a 0 to 10% EtOAc gradient in hexane to give the aldehyde (950 mg): H NMR (400 MHz, CDCl 3 ) 6 11.61 (s, 1H), 9.91 (s, 1H), 7.79 (d, J= 2.5 Hz, 1H), 7.75 (d, J = 2.6 Hz, 1H); EIMS m/z 224. [0053] Example 3. Preparation of 3-chloro-2-hydroxy-5 trifluoromethylbenzaldeyde CI OH F N CHO F F 3-Chloro-2-fluoro-5-trifluoromethylbenzaldehyde (5.0 g, 22 mmol) was dissolved in dry

CH

3 0H (50 mL), treated with 25% sodium methoxide solution (30 mL) and heated to reflux for 2 h. After cooling, the volatiles were removed by evaporation and the residue was taken up in H 2 0 (20 mL) plus Et 2 0 (80 mL). The aqueous phase was extracted with Et 2 0 (50 mL), and the combined organic phases were washed with satd NaCl solution (15 mL), dried (Na 2

SO

4 ) and evaporated. The residue was dissolved in dry CH 2 Cl 2 (50 mL), cooled to -78 'C and treated with BBr 3 (1 M solution in CH 2 Cl 2 ; 25 mL, 25 mmol). After warming to 25 'C, the mixture was stirred for 21 h, cooled to -40 'C and quenched by addition of H 2 0 (30 mL). After warming, the aqueous phase was extracted with CH 2 Cl 2 (30 mL), and the combined organic phases were washed with satd NaCl solution (30 mL), dried (Na 2

SO

4 ) and 10 WO 2012/024494 PCT/US2011/048260 evaporated. The residue was purified by silica gel chromatography with a 0-20% EtOAc gradient in hexane to give the purified aldehyde (2.7 g): 1 H NMR (400 MHz, CDCl 3 ) 6 11.81 (s, 1H), 9.96 (s, 1H), 7.87 (d, J= 2.1 Hz, 1H), 7.84-7.77 (m, 1H); EIMS m/z 224. [0054] Example 4. Preparation of 3-chloro-2-hydroxy-6-trifluoromethyl benzaldehyde CI OH CHO

CF

3 3-Chloro-2-fluoro-6-trifluoromethylbenzaldehyde (1.0 g, 4.4 mmol) was dissolved in dry

CH

3 0H (10 mL), treated with 30% sodium methoxide solution in CH 3 0H (7.9 g, 44 mmol) and heated at reflux for 1 h. After cooling, the mixture was diluted with H 2 0 (15 mL) and extracted with Et 2 0 (30 mL). The combined organic extracts were washed with satd NaCl solution (10 mL), dried (Na 2 SO4), and evaporated. The residue was purified by silica gel chromatography with a 0-10% EtOAc gradient in hexane to give the anisole intermediate (1.0 g). This material was dissolved in dry CH 2 Cl 2 (15 mL), cooled to -78 'C, treated with BBr 3 (1 M in CH 2 Cl 2 ; 5.0 mL, 5 mmol), allowed to warm to 25 'C and stirred for 20 h. The reaction was cooled in ice and quenched by addition of H 2 0 (10 mL). The separated organic phase was washed with satd NaCl solution (10 mL), dried (Na 2

SO

4 ) and evaporated. The residue was purified by silica gel chromatography with a 0-10% EtOAc gradient in hexane to give the aldehyde (980 mg): 1 H NMR (400 MHz, CDCl 3 ) 6 12.78 (s, 1H), 10.28 (s, 1H), 7.71 (d, J = 8.2 Hz, 1H), 7.27 (d, J= 8.5 Hz, 1H); EIMS m/z 224. [0055] Example 5. Preparation of 3,4-dichloro-2-hydroxybenzaldehyde CI CI OH CHO 3,4-Dichloro-2-hydroxybenzaldehyde was prepared from commercially available starting materials as described in Gu et al., J. Med. Chem. 2000, 43, 4868-4876, which is expressly incorporated by reference herein. [0056] Example 6. Preparation of 3,6-dichloro-2-hydroxybenzaldehyde CI OH CHO CI 11 WO 2012/024494 PCT/US2011/048260 3,6-Dichloro-2-hydroxybenzaldehyde was prepared from commercially available starting materials as described in Rafferty et al., PCT Int. Appl. WO 2008121602 Al, which is expressly incorporated by reference herein. [0057] Example 7. Preparation of 2-hydroxy-5-trifluoromethylbenzaldehyde OH

F

3 C CHO 2-Hydroxy-5-trifluoromethylbenzaldehyde was prepared from commercially available starting materials as described in Bonnert et al., PCT Int. Appl. WO 2006056752 Al, which is expressly incorporated by reference herein. [0058] Example 8. Preparation of 4-chlorobenzoic acid [1-(2-chloro-6 hydroxyphenyl)-methylidene]-hydrazide (Compound 70) OH 0 CI N' CI A suspension of 2-chloro-6-hydroxybenzaldehyde (0.10 g, 0.64 mmol) and 4-chlorobenzoic acid hydrazide (0.10 g, 0.61 mmol) in EtOH (3 mL) was agitated and heated to 60 'C for 16 h. The reaction mixture was cooled to room temperature and H 2 0 (1-2 mL) was added portionwise to precipitate the product. The solid was collected via suction filtration and rinsed with ethanol to furnish the title compound as an off-white solid (0.18 g, 95%): mp 273-277 'C; 'H NMR (400 MHz, DMSO-d 6 ) 6 12.53 (s, 1H), 12.51 (s, 1H), 9.04 (s, 1H), 8.03 - 7.95 (m, 2H), 7.71 - 7.63 (m, 2H), 7.34 (t, J= 8.2 Hz, 1H), 7.07 (dd, J= 7.9, 0.9 Hz, 1H), 6.96 (d, J= 8.2 Hz, 1H); ESIMS m/z 309 ([M+H]*), 307 ([M-H]-). [0059] Example 9. Preparation of 3-methyl-benzoic acid [1-(3,5-dichloro-2 hydroxyphenyl)-ethylidene]-hydrazide (Compound 25) CI OH 0 CI N N A suspension of 1-(3,5-dichloro-2-hydroxyphenyl)-ethanone (0.100 g, 0.49 mmol), 3 methylbenzoic acid hydrazide (0.069 g, 0.46 mmol), and glacial acetic acid (0.3 mL) in EtOH (3 mL) was heated to 60 'C for 4 h The reaction mixture was cooled to room temperature to precipitate the product. The solid was collected via suction filtration and rinsed with H 2 0 to furnish the title material as a yellow solid (0.144 g, 93%): mp 221-223 'C; 1 H NMR (400 12 WO 2012/024494 PCT/US2011/048260 MHz, DMSO) 6 14.48 (s, 1H), 11.54 (s, 1H), 7.79 - 7.72 (m, 2H), 7.68 (d, J= 2.5 Hz, 1H), 7.64 (d, J = 2.4 Hz, 1H), 7.47 - 7.42 (m, 2H), 2.52 (s, 3H), 2.41 (s, 3H); ESIMS m/z 337 ([M+H]*), 335 ([M-H]-). Table 1. Structures of Exemplified Compounds Compound Structure Number CI OH C OH 0 1 H CH OH 2 H N 3HC HI CI OH O Br 5H CI 13 WO 2012/024494 PCT/US2011/048260 Compound Structure Number CI OH 7 -CN 0 OH CI N H CI :-' OH 0 8 CIN CI 10 C N H I 3C I CI N CH 14: OH 10 N,. HI Br 11 6C0 Br '.~ N H I CI 12 I N, CI 13 O Cl H OH, 14 WO 2012/024494 PCT/US2011/048260 Compound Structure Number Br 14 ci HN H Br 15 ~ OH H Br OH 16 -l C-' 0 QOH H Br O H 17 Z:-B N, Br H Br OH H 2 B CHr OH Br-& :N N Br H 20 0 Br N, N H 'J~ CH, 15 WO 2012/024494 PCT/US2011/048260 Compound Structure Number CI CI OHO 0 21 Cl-,C N' H CI O N N H 27 (t H O OH C N 28 FN F H CI H 0 FE, H CI OH 25 0 26 N. O

CH

3 H I CH OH 26 F - N N F F F F1 WO 2012/024494 PCT/US2011/048260 70130 Compound Structure Number CI 29 F H- NF F F C 33 F F& N0O OH 30 FEI0O F

HI

CI O OH 31 F F, N 1.1 1~. F F 32 OH C F1 HI F 33 LOH0 34 NH OH H 34 LOH0 H0j~~ - .~N17 WO 2012/024494 PCT/US2011/048260 Compound Structure Number 36 F OH O OH N F F OH 37 OH O OH N F 38 CI H CH, CI F F Ci~i OH I 37 0 OH 3N H H II CI O 390 CN, C3 H CI CIj:,CF OH 40 ~ N HH, CI 42 H OH Cl H 18 WO 2012/024494 PCT/US2011/048260 Compound Structure Number CI OHO 43 C N l HI CI 44 OH C N, CI l HI CI OHO 45 N C N 47 HO C N 46 H 0 OH C N H 47 OH 0 49 N, N 19 WO 2012/024494 PCT/US2011/048260 Compound Structure Number CI 50 OH O N CH3 Nd HNjt( CH, H CH 51 I H 0 N 6 N C H CI 52 CHN0 N NCI C5 0H 55 OH 0 OH NN CI H 54 0 O - NUNab I OH0 WO 2012/024494 PCT/US2011/048260 Compound Structure Number CI OH 57 N,0 OH F F C CI OH 00 OH 58 Br N 63H CI N CH OH1 59 N.. N HI CH, B HrB 61 N B r - N OH HI F 62 (; CH Njja l CI1 63 JC : Ni N H 21 WO 2012/024494 PCT/US2011/048260 Compound Structure Number CI 64 0i~ N~ CI HNCI CI 65 CI NC 66 N- C CI H 68F F 66 OH 0 69 N F H CI O OH OH ;7 H 0 68 F - _ N,1 7 F HI 20 69 _ , F l HI 70 _ N 'N 71 F

-

N-H 11 22 WO 2012/024494 PCT/US2011/048260 Table 2. Analytical Data for Compounds in Table 1 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) ) (-) unless otherwise stated 1 309 307 (300 MHz, CDCl 3 ) 12.73 (s, 1H), 9.00 (s, 1H), 7.90 - 7.82 (m, 2H), 2 185-187 255 253 7.64 - 7.57 (m, 1H), 7.55 - 7.47 (m, 3H), 7.35 - 7.28 (m, 1H), 7.09 - 7.00 (m, 1H), 6.94 - 6.87 (m, 1H), 2.42 (s, 3H) 3 275 273 4 327 325 14.32 (s, 1H), 11.91 (s, 1H), 7.76 (dd, J= 7.8, 1.1 Hz, 1H), 7.67 (d, J 5 220-222 403 401 = 2.5 Hz, 1H), 7.65 (d, J= 2.4 Hz, 1H), 7.61 (dd, J = 7.4, 1.8 Hz, 1H), 7.55 - 7.45 (m, 2H), 2.45 (s, 3H) 6 321 (M+Na) 7 323 8 343 341 23 WO 2012/024494 PCT/US2011/048260 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) (-) unless otherwise stated 9 385 12.70 (s, 1H), 12.60 (s, 1H), 8.63 10 215-217 359 (s, 1H), 8.60 (s, 1H), 8.13 - 8.07 (m, 2H), 8.06 - 8.00 (m, 2H), 7.73 - 7.62 (m, 4H) 11 421 397 (M+Na) 14.41 (s, 1H), 11.65 (s, 1H), 7.99 12 260-262 355 (d, J = 8.5 Hz, 2H), 7.68 (d, J = 2.5 Hz, 1H), 7.66 - 7.60 (m, 3H), 2.68 - 2.32 (m, 3H) 14.27 (s, 1H), 11.44 (s, 1H), 7.61 13 199-204 338 336 (dd, J= 10.6, 2.5 Hz, 2H), 7.36 7.32 (m, 4H), 7.30 - 7.23 (m, 1H), 3.75 (s, 2H), 2.44 (s, 3H) 14 351 (M+Na) 15 411 (M+Na) 16 367 24 WO 2012/024494 PCT/US2011/048260 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) ) (-) unless otherwise stated 17 456 (M+Na) 18 367 19 429 427 20 363 361 12.88 (s, 1H), 12.46 (s, 1H), 8.61 21 249-252 289 287 (s, 1H), 8.04 - 7.89 (m, 2H), 7.60 (ddd, J= 12.6, 11.5, 6.4 Hz, 4H), 7.23 (d, J= 8.4 Hz, 1H) 12.70 (s, 1H), 12.38 (s, 1H), 8.60 (s, 1H), 8.32 - 8.23 (m, 2H), 8.02 22 165-169 375 (d, J = 7.8 Hz, 1H), 7.83 (t, J = 7.8 Hz, 1H), 7.68 (dd, J= 24.6, 2.5 Hz, 2H) 15.17 (s, 1H), 11.70 (s, 1H), 7.99 (d, J= 8.5 Hz, 3H), 7.94 (dd, J= 23 241-251 334 332 8.1, 1.4 Hz, 1H), 7.65 (d, J= 8.5 Hz, 2H), 7.06 (t, J = 8.0 Hz, 1H), 2.56 (s, 3H) 13.24 (s, 1H), 12.64 (s, 1H), 8.69 24 216-222 343 341 (s, 1H), 8.04 - 7.88 (m, 4H), 7.62 (dt, J= 31.2, 7.3 Hz, 3H) 25 WO 2012/024494 PCT/US2011/048260 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) (-) unless otherwise stated 14.48 (s, 1H), 11.54 (s, 1H), 7.79 7.72 (m, 2H), 7.68 (d, J= 2.5 Hz, 25 221-223 337 335 1H), 7.64 (d, J= 2.4 Hz, 1H), 7.47 - 7.42 (m, 2H), 2.52 (s, 3H), 2.41 (s, 3H) 26 262-264 337 335 13.15 (s, 1H), 12.64 (s, 1H), 8.61 27 225-226 343 341 (s, 1H), 8.04 - 7.90 (m, 3H), 7.75 7.52 (m, 4H) 13.15(s, 1H), 12.69 (s, 1H), 8.69 (s, 1H), 8.03 (s, 1H), 7.91 (d, J= 1.8 28 210-212 361 359 Hz, 1H), 7.80 (dd, J= 19.3, 8.7 Hz, 2H), 7.64 (d, J= 5.8 Hz, 1H), 7.52 (d, J= 1.9 Hz, 1H) 13.4 (s, 1H), 12.66 (s, 1H), 8.68 (s, 29 200-202 361 359 1H), 8.11 - 7.98 (m, 2H), 7.90 (d, J = 2.1 Hz, 1H), 7.43 (t, J= 8.8 Hz, 2H), 7.35 - 7.24 (m, 1H) 13.11 (s, 1H), 12.35 (s, 1H), 11.45 (s, 1H), 8.73 (s, 1H), 7.98 (d, J= 30 258-260 359 357 1.7 Hz, 1H), 7.95 - 7.83 (m, 2H), 7.54 - 7.40 (m, 1H), 7.00 (dd, J= 14.0, 7.4 Hz, 2H) 13.09 (s, 1H), 12.37 (s, 1H), 11.63 (s, 1H), 8.72 (s, 1H), 7.97 (d, J= 31 255-257 373 371 1.7 Hz, 1H), 7.90 (d, J= 1.8 Hz, 1H), 7.81 (d, J= 8.5 Hz, 1H), 6.81 (d, J= 8.5 Hz, 2H), 2.32 (d, J= 6.7 Hz, 3H) 13.85 (s, 1H), 12.84 (s, 1H), 8.71 32 233-236 411 409 (s, 1H), 8.36 (s, 1H), 8.00 (d, J= 8.5 Hz, 2H), 7.94 (s, 1H), 7.67 (d, J = 8.5 Hz, 2H) 26 WO 2012/024494 PCT/US2011/048260 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) (-) unless otherwise stated 13.03 (s, 1H), 12.44 (s, 1H), 8.64 (s, 1H), 7.82 - 7.72 (m, 3H), 7.67 33 188-190 323 321 (d, J = 7.3 Hz, 1H), 7.46 (d, J = 5.0 Hz, 2H), 7.11 (t, J= 7.8 Hz, 1H), 2.41 (s, 3H) (300 MHz) 12.92 (s, 1H), 12.54 (s, 1H), 8.65 (s, 1H), 7.86 - 7.73 (m, 34 176-180 327 325 3H), 7.72 - 7.59 (m, 2H), 7.51 (td, J= 8.5, 2.2 Hz, 1H), 7.12 (t, J= 7.7 Hz, 1H) 12.96 (s, 1H), 12.55 (s, 1H), 8.64 (s, 1H), 7.99 (d, J= 8.6 Hz, 2H), 35 215-216 343 341 7.81 (d, J= 7.5 Hz, 1H), 7.71 7.63 (m, 3H), 7.11 (t, J= 7.7 Hz, 1H) 12.95 (s, 1H), 12.28 (s, 1H), 11.56 (s, 1H), 8.71 (s, 1H), 7.88 (dd, J= 36 230-234 325 323 7.9, 1.6 Hz, 1H), 7.79 (d, J = 7.6 Hz, 1H), 7.68 (d, J= 7.1 Hz, 1H), 7.52 - 7.44 (m, 1H), 7.12 (t, J= 7.8 Hz, 1H), 7.05 - 6.96 (m, 2H) 12.95 (s, 1H), 12.26 (s, 1H), 11.67 (s, 1H), 8.70 (s, 1H), 7.80 (t, J= 9.1 37 248-253 339 337 Hz, 2H), 7.67 (d, J = 7.4 Hz, 1H), 7.11 (t, J= 7.8 Hz, 1H), 6.83 (d, J= 7.4 Hz, 2H), 2.31 (s, 3H) 12.89 (s, 1H), 12.41 (s, 1H), 8.60 (s, 1H), 7.79 - 7.70 (m, 2H), 7.54 38 212-214 323 321 (d, J = 8.5 Hz, 1H), 7.45 (d, J = 4.4 Hz, 2H), 7.24 (d, J = 8.4 Hz, 1H), 2.41 (s, 3H) 12.75 (s, 1H), 12.50 (s, 1H), 8.61 (s, 1H), 7.81 (d, J= 7.8 Hz, 1H), 39 215-218 327 325 7.79 - 7.73 (m, 1H), 7.67 - 7.59 (m, 1H), 7.58 (d, J= 8.5 Hz, 1H), 7.51 (td, J= 8.4, 2.4 Hz, 1H), 7.24 (d, J= 8.4 Hz, 1H) 12.73 (s, 1H), 12.52 (s, 1H), 8.60 (s, 1H), 8.00 (t, J= 1.8 Hz, 1H), 40 219-222 343 341 7.92 (d, J = 7.8 Hz, 1H), 7.74 7.69 (m, 1H), 7.61 (t, J= 7.9 Hz, 1H), 7.57 (d, J= 8.5 Hz, 1H), 7.24 (d, J= 8.4 Hz, 1H) 27 WO 2012/024494 PCT/US2011/048260 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) (-) unless otherwise stated 12.79 (s, 1H), 12.24 (s, 1H), 11.69 (s, 1H), 8.66 (s, 1H), 7.80 (d, J= 41 271-278 339 337 8.6 Hz, 1H), 7.54 (d, J= 8.5 Hz, 1H), 7.24 (d, J= 8.4 Hz, 1H), 6.84 - 6.79 (m, 2H), 2.31 (s, 3H) 13.46 (s, 1H), 12.57 (s, 1H), 9.01 (s, 1H), 7.82 - 7.75 (m, 2H), 7.53 42 214-216 323 (d, J = 8.6 Hz, 1H), 7.49 - 7.45 (m, 2H), 7.10 (d, J= 8.6 Hz, 1H), 2.42 (s, 3H) 13.41 (s, 1H), 12.62 (s, 1H), 9.00 43 264-267 327 325 (s, 1H), 8.10 - 8.01 (m, 2H), 7.53 (d, J = 8.6 Hz, 1H), 7.44 (t, J = 8.8 Hz, 2H), 7.10 (d, J= 8.6 Hz, 1H) 13.34 (s, 1H), 12.67 (s, 1H), 9.00 (s, 1H), 8.04 - 8.00 (m, 1H), 7.94 44 232-234 343 341 (d, J = 7.8 Hz, 1H), 7.74 (d, J = 9.0 Hz, 1H), 7.63 (t, J= 7.9 Hz, 1H), 7.54 (d, J= 8.6 Hz, 1H), 7.10 (d, J = 8.6 Hz, 1H) 13.38 (s, 1H), 12.66 (s, 1H), 9.00 (s, 1H), 8.00 (d, J= 8.5 Hz, 2H), 45 271-273 343 7.67 (d, J= 8.5 Hz, 2H), 7.53 (d, J = 8.6 Hz, 1H), 7.10 (d, J= 8.6 Hz, 1H) 13.43 (s, 1H), 12.46 (s, 1H), 11.48 (s, 1H), 9.06 (s, 1H), 7.86 (dd, J= 46 260-262 325 323 7.9, 1.6 Hz, 1H), 7.54 (d, J = 8.6 Hz, 1H), 7.52 - 7.45 (m, 1H), 7.10 (d, J = 8.6 Hz, 1H), 7.05 - 6.96 (m, 2H) 13.08 (s, 1H), 12.74 (s, 1H), 8.60 47 228-229 377 375 (s, 1H), 8.05 - 7.97 (m, 2H), 7.93 (d, J = 7.8 Hz, 1H), 7.72 (t, J = 5.3 Hz, 2H), 7.62 (t, J= 7.9 Hz, 1H) 13.10 (s, 1H), 12.43 (s, 1H), 11.51 (s, 1H), 8.66 (s, 1H), 7.97 (d, J= 48 258-260 359 357 2.5 Hz, 1H), 7.87 (dd, J= 7.9, 1.5 Hz, 1H), 7.71 (d, J= 2.5 Hz, 1H), 7.55 - 7.39 (m, 1H), 7.06 - 6.91 (m, 2H) 28 WO 2012/024494 PCT/US2011/048260 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) (-) unless otherwise stated 12.73 (s, 1H), 12.44 (s, 1H), 9.06 (s, 1H), 8.46 - 8.38 (m, 2H), 8.25 49 >300 320 318 8.16 (m, 2H), 7.36 (t, J= 8.2 Hz, 1H), 7.08 (dd, J = 8.0, 0.9 Hz, 1H), 6.97 (d, J= 8.1 Hz, 1H) (400 MHz, CDCl 3 ) 8.59 (d, J= 12.0 Hz, 1H), 7.76 (d, J = 10.0 Hz, 2H), 50 166-169 289 287 7.47 (dd, J= 15.6, 6.2 Hz, 4H), 6.97 (t, J= 7.8 Hz, 1H), 2.39 (d, J= 12.0 Hz, 3H) (400 MHz, CDCl 3 ) 8.61 (s, 1H), 8.00 (t, J= 1.7 Hz, 1H), 7.95 - 7.68 51 174-178 309 307 (m, 2H), 7.61 (t, J= 7.9 Hz, 1H), 7.55 - 7.47 (m, 2H), 6.98 (t, J = 7.8 Hz, 1H) 12.20 (s, 1H), 11.50 (s, 1H), 8.64 52 257-260 309 307 (s, 1H), 7.96 (d, J= 8.5 Hz, 2H), 7.67-7.60 (m, 3H), 7.03-6.94 (m, 2H) (400 MHz, CDCl 3 ) 8.64 (s, 1H), 53 284-287 320 318 8.41 (d, J= 8.5 Hz, 2H), 8.19 (d, J = 8.6 Hz, 2H), 7.56 - 7.47 (m, 2H), 6.98 (t, J= 7.8 Hz, 1H) (400 MHz, CDCl 3 ) 8.66 (s, 1H), 54 266-268 291 289 7.87 (dd, J= 7.9, 1.6 Hz, 1H), 7.54 - 7.42 (m, 3H), 6.98 (ddd, J= 7.8, 5.6, 3.9 Hz, 3H) (400 MHz, CDCl 3 ) 8.66 (s, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.49 (dd, J 55 258-266 305 303 = 7.9, 2.1 Hz, 2H), 6.98 (t, J= 7.8 Hz, 1H), 6.81 (d, J= 7.3 Hz, 2H), 2.31 (s, 3H) 13.76 (s, 1H), 12.73 (s, 1H), 8.92 (s, 1H), 8.00 (d, J= 8.6 Hz, 2H), 56 236-238 377 375 7.72 (d, J = 8.3 Hz, 1H), 7.67 (d, J = 8.6 Hz, 2H), 7.36 (d, J= 8.4 Hz, 1H) 29 WO 2012/024494 PCT/US2011/048260 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) (-) unless otherwise stated 13.82 (s, 1H), 12.57 (s, 1H), 11.28 (s, 1H), 8.98 (d, J= 1.3 Hz, 1H), 57 278-280 358 356 7.84 (dd, J= 7.9, 1.5 Hz, 1H), 7.73 (d, J = 8.3 Hz, 1H), 7.53 - 7.45 (m, 1H), 7.36 (d, J= 8.4 Hz, 1H), 7.01 (dd, J= 13.2, 7.7 Hz, 2H) 14.55, 11.80, 11.69, 11.46, 9.82, 9.72 (6s, 3H), 7.64 - 7.50 (m, 2H), 58 245 352 351 7.16 (t, J= 7.9 Hz, 1H), 6.77 - 6.71 (m, 2H), 2.40, 2.38 (2s, 3H), 2.22, 2.14 (2s, 3H) Note: rotational isomers 14.38 (s, 1H), 11.40 (s, 1H), 7.87 (d, J = 8.0 Hz, 2H), 7.64 (dd, J = 59 195-200 303 301 8.1, 1.5 Hz, 1H), 7.49 (dd, J= 7.9, 1.3 Hz, 1H), 7.37 (d, J= 8.0 Hz, 2H), 6.92 (t, J= 8.0 Hz, 1H), 2.51 (s, 3H), 2.40 (s, 3H) 15.16 (s, 1H), 11.70 (s, 1H), 8.02 60 244-252 379 7.87 (m, 4H), 7.79 (d, J= 8.5 Hz, 2H), 7.06 (t, J = 8.0 Hz, 1H), 2.56 (s, 3H) 61 242-245 414 13.48 (s, 1H), 11.59 (s, 1H), 7.98 62 276-280 325 323 (d, J = 8.5 Hz, 2H), 7.64 (d, J = 8.5 Hz, 2H), 7.43 - 7.36 (m, 2H), 2.50 (s, 3H) 63 323 321 64 343 341 30 WO 2012/024494 PCT/US2011/048260 Compound a ESIMS ESIMS 'H NMR (400 MHz, DMSO-d 6 ) 6, Number mp (C) (-) unless otherwise stated 14.38 (s, 1H), 11.68 (s, 1H), 8.02 (s, 1H), 7.91 (d, J= 7.7 Hz, 1H), 65 245-246 357 355 7.72 (d, J= 9.1 Hz, 1H), 7.70 (d, J = 2.5 Hz, 1H), 7.65 (d, J= 2.4 Hz, 1H), 7.60 (t, J = 7.9 Hz, 1H), 2.53 (s, 3H) 13.13 (s, 1H), 12.72 (s, 1H), 8.60 66 232-234 377 375 (s, 1H), 8.00 (dd, J= 10.9, 5.5 Hz, 3H), 7.71 (d, J= 2.4 Hz, 1H), 7.66 (d, J= 8.6 Hz, 2H) 67 12.25 (s, 1H), 11.84 (s, 1H), 8.73 68 214-217 309 307 (s, 1H), 8.01 (s, 1H), 7.95 (d, J= 7.9 Hz, 2H), 7.59 (dt, J= 30.0, 7.5 Hz, 4H), 7.12 (d, J= 8.6 Hz, 1H) 12.52 (s, 1H), 12.48 (s, 1H), 9.04 (s, 1H), 7.83 (d, J= 7.8 Hz, 1H), 7.80 - 7.75 (m, 1H), 7.68 - 7.60 69 257-259 293 291 (m, 1H), 7.52 (td, J= 8.5, 2.4 Hz, 1H), 7.35 (t, J = 8.2 Hz, 1H), 7.07 (d, J = 7.9 Hz, 1H), 6.97 (d, J = 8.3 Hz, 1H) 12.53 (s, 1H), 12.51 (s, 1H), 9.04 (s, 1H), 8.03 - 7.95 (m, 2H), 7.71 70 273-277 309 307 7.63 (m, 2H), 7.34 (t, J= 8.2 Hz, 1H), 7.07 (dd, J = 7.9, 0.9 Hz, 1H), 6.96 (d, J = 8.2 Hz, 1H) 12.15 (s, 1H), 11.78 (s, 2H), 8.75 (s, 1H), 8.01 (d, J= 1.8 Hz, 1H), 71 277-281 325 323 7.89 (dd, J= 7.8, 1.2 Hz, 1H), 7.65 (dd, J= 8.6, 2.1 Hz, 1H), 7.50 7.42 (m, 1H), 7.13 (d, J= 8.6 Hz, 1H), 7.03 - 6.94 (m, 2H) [0060] Example 10. Effect of copper on activity of hydrazones towards Chlamydomonas reinhardtii In vitro toxicity assays against C. reinhardtii CC-1928 (wild type mt+ 137c green in dark subclone) obtained from the Chlamydomonas Resource Center at Duke University were 31 WO 2012/024494 PCT/US2011/048260 conducted using TAP liquid growth medium described by Gorman and Levine Proc. NatL. Acad. Sci. USA 1965, 54, 1665-1669 except that CuSO 4 -5H 2 0, normally included as a component of Hutner's trace element stock solution (Hutner et al. Proc. Am. Philos. Soc. 1950, 94, 152-170), was omitted. The TAP medium lacking Cu (TAP Cu minus) was prepared by adding 400 mg NH 4 Cl, 10 mL of 100X Tris-Acetate (242 g Tris-base and 100 mL glacial acetic acid in 1000 mL milli Q water), and 8.3 mL of 120.5X phosphate buffer (14.34 g K 2

HPO

4 and 7.26 g KH 2

PO

4 in 1000 mL milli Q water) to milli Q water to make a total volume of 1 liter. The medium was treated with 0.5 g Chelex 100 resin (Bio-Rad Analytical grade, 50-100 mesh, sodium form, cat# 142-2822) by stirring at room temperature for 1 hour. 900 mL were decanted and 45.9 mg of CaCl 2 -2H 2 0, 90 mg of MgSO 4 -7H 2 0 and 0.9 mL Hutner's trace elements lacking CuSO 4 -5H 2 0 were added and the solution was filter sterilized. Medium containing copper (TAP Cu plus) was prepared by adding CuCl 2 -2H 2 0 to the TAP Cu minus medium at 20 pM. Test compounds were dissolved in dimethylsulfoxide (DMSO) at 2 mg/mL then dilutions in TAP Cu minus and TAP Cu plus media were prepared in sterilized microcentrifuge tubes at 2 pg/mL. 100 pL aliquots of diluted compound were dispensed into flat-bottomed 96-well microtiter plates. [0061] C. reinhardtii was maintained under 12 h of light on TAP agar plates at 18 'C. A loopful of cells was added to 50 mL of TAP Cu minus medium and the culture incubated with shaking at 200 rpm at 27 C for 24 h. The culture (10 to 20 mL aliquot) was centrifuged in a bench centrifuge at 2000 rpm for 2 min. The resulting cell pellet was resuspended in 5 mL sterile milli Q water (which had been treated with Chelex 100 resin using 0.5 g resin per liter of water by stirring at room temperature for 1 h), and recentrifuged. The cells were resuspended in TAP Cu minus medium, and the suspension adjusted to 2 x 10 5 spores per mL. Microtiter plates containing compounds as described above were inoculated with 100 ptL of this cell suspension and the plates incubated at 27 'C for 48 h before assessing algal growth by measuring light scattering in a NepheloStar plate-reader. Growth inhibition was determined by comparing growth in the presence of test compound with growth in control wells lacking test compound. [0062] Results for growth inhibition by test compounds at 1 ptg/mL in TAP Cu plus medium (% Inhn. Plus Copper Observed) were compared with predicted values (% Inhn. Plus Copper Predicted) that were calculated using the formula set forth by S. R. Colby in Weeds 1967, 15, 20-22 based on results obtained for the same compounds in TAP Cu minus medium (% Inhn. Minus Copper Observed). Copper chloride alone caused a slight enhancement of 32 WO 2012/024494 PCT/US2011/048260 growth based on comparison of growth in TAP Cu minus and TAP Cu plus media without any test compound across experiments. Data are presented in Table 3. Results illustrate that hydrazones and copper produce a synergistic algicidal effect towards C. reinhardtii. Table 3. Growth Inhibition of C. reinhardtii %Inhn. % Inhn. % Inhn. Minus Plus Plus Compound Copper Copper Copper Number Observed Observed Predicted 1 16 96 6 2 10 95 -1 3 25 62 16 4 12 92 2 5 26 95 17 6 13 98 3 7 29 94 20 8 15 92 5 9 7 90 -3 10 24 93 15 11 31 96 22 12 4 96 -7 13 17 96 7 14 6 97 -5 15 11 97 1 16 -3 95 -15 17 32 83 24 18 13 95 3 19 12 52 2 20 -6 94 -19 21 6 94 -5 22 -14 96 -27 23 -37 89 -53 24 -2 90 -14 25 33 95 26 26 5 94 -6 27 30 96 21 28 12 97 2 29 16 96 7 30 29 92 20 31 -1 88 -13 32 21 97 12 33 21 97 11 33 WO 2012/024494 PCT/US2011/048260 %Inhn. % Inhn. % Inhn. Minus Plus Plus Compound Copper Copper Copper Number Observed Observed Predicted 34 8 97 -2 35 9 97 -1 36 7 96 -4 37 12 97 1 38 15 94 5 39 8 91 -3 40 22 96 12 41 28 96 19 42 10 96 0 43 9 93 -2 44 11 94 0 45 9 96 -2 46 0 92 -12 47 -5 77 -18 48 18 92 8 49 33 93 25 50 6 97 -5 51 -13 96 -26 52 -4 88 -16 53 8 88 -3 54 -8 75 -21 55 -3 97 -15 56 11 97 0 57 -11 95 -24 58 3 93 -8 59 -6 92 -19 60 -50 89 -68 61 -56 94 -75 62 0 95 -12 63 -18 93 -32 64 -5 97 -18 65 -5 97 -18 66 -4 97 -17 CuCl 2 , 10 M -11.7 ± 6.0 34 WO 2012/024494 PCT/US2011/048260 [0063] Example 11. Effect of copper on activity of hydrazones towards Synechocystis In vitro toxicity assays against the cyanobacterium (blue-green alga) Synechocystis sp. strain 6803 were conducted using BG- 11 medium except that copper micronutrient, normally included as CuSO 4 -5H 2 0, was omitted. The medium, termed BG-1 1 Cu minus, was prepared by dissolving 1.5 g NaNO 3 , 40 mg K 2

HPO

4 , 6 mg citric acid, 1 mg Na 2 EDTA and 20 mg Na 2

CO

3 in 1 liter of milli Q water and treating the solution with 0.5 g Chelex 100 resin (Bio Rad Analytical grade, 50-100 mesh, sodium form, cat# 142-2822) by stirring at room temperature for 1 h. 900 mL were decanted and 67.5 mg MgSO 4 -7H 2 0, 32.4 mg CaCl 2 -7H 2 0, 5.4 mg ferric ammonium citrate and 0.9 mL trace metal mix A5 without copper (2.86 g H 3 B0 3 , 1.81 g MnCl 2 -4H 2 0, 222 mg ZnSO 4 -7H 2 0, 390 mg Na 2 MoO 4 -2H 2 0 and Co(N0 3 ) 2-6H 2 0 in 1 L Milli Q water) was added. The pH was adjusted to 7.1 and the medium was filter-sterilized. Medium containing copper (BG- 11 Cu plus) was prepared by adding CuCl 2 -2H 2 0 to the BG-1 1 Cu minus medium at 0.2 pM. Test compounds were dissolved in dimethylsulfoxide (DMSO) at 2 mg/mL then appropriate dilutions in BG- 11 Cu minus and BG-11 Cu plus media were prepared in sterilized microcentrifuge tubes. 100 IL aliquots were dispensed into flat-bottomed 96-well microtiter plates. [0064] Synechocystis was maintained under 24 h of light on BG- 11 agar plates at 27 'C. A loopful of cells was added to 50 mL of BG- 11 Cu minus growth medium and the culture was incubated with shaking at 200 rpm at 27 'C for 24 h. The culture (10 to 20 mL aliquot) was centrifuged in a bench centrifuge at 2400 rpm for 2 min. The resulting cell pellet was resuspended in 5 mL sterile milli Q water (which had been treated with Chelex 100 resin using 0.5 g resin per liter of water by stirring at room temperature for 1 h), and recentrifuged. The cells were resuspended in BG-11 Cu minus medium, and the suspension adjusted to 4 x 10 5 spores per mL. Microtiter plates containing compounds as described above were inoculated with 100 ptL of this cell suspension and the plates incubated at 27 OC for 72 h before assessing algal growth by measuring light scattering in a NepheloStar plate reader. Growth inhibition was determined by comparing growth in the presence of test compound with growth in control wells lacking test compound. [0065] Results for growth inhibition by test compounds in BG- 11 Cu plus medium (% Inhn. Plus Copper Observed) were compared with predicted values (% Inhn. Plus Copper Predicted) that were calculated using the formula set forth by S. R. Colby in Weeds 1967, 15, 20-22 based on-results obtained for the same compounds in BG- 11 Cu minus medium (% 35 WO 2012/024494 PCT/US2011/048260 Inhn. Minus Copper Observed) and the inhibition attributed to copper chloride alone, as determined by comparing growth in copper-minus and copper-plus media without any test compound across experiments. Data are presented in Table 4. Results illustrate that hydrazones and copper produce a synergistic algicidal effect towards Synechocystis. Table 4. Growth Inhibition of Synechocystis Test %Inhn. % Inhn. % Inhn. Conen. Minus Plus Plus Compound Cg/mL Copper Copper Copper Number Observed Observed Predicted 1 0.05 29 91 34 2 0.05 -8 97 -1 3 0.05 -17 96 -9 4 0.05 -14 96 -6 5 0.05 -10 91 -2 6 0.05 19 99 25 7 0.05 22 96 28 8 0.05 62 92 65 9 0.00625 15 74 21 10 0.05 44 95 48 11 0.05 50 97 54 12 0.0125 21 82 27 13 0.05 22 96 28 14 0.05 50 97 54 15 0.00625 15 74 21 16 0.05 -5 94 3 17 0.00625 59 72 62 18 0.0125 47 86 51 19 0.05 -18 43 -9 20 0.05 9 86 15 21 0.0125 39 96 43 22 0.05 48 94 52 23 0.0125 27 92 32 24 0.05 45 93 49 25 0.05 18 92 23 26 0.05 54 95 57 27 0.05 31 95 36 28 0.05 13 96 19 29 0.05 11 96 18 30 0.05 0 92 7 31 0.05 38 92 42 32 0.05 26 91 31 36 WO 2012/024494 PCT/US2011/048260 Test %Inhn. % Inhn. % Inhn. Conen. Minus Plus Plus Compound pg/mL Copper Copper Copper Number Observed Observed Predicted 33 0.05 29 97 34 34 0.05 34 91 39 35 0.05 26 91 31 36 0.0125 28 87 33 37 0.0125 44 88 48 38 0.05 49 93 53 39 0.05 31 92 36 40 0.00625 4 64 11 41 0.0125 26 93 31 42 0.05 10 94 16 43 0.05 23 95 28 44 0.05 61 93 63 45 0.00625 33 78 38 46 0.0125 37 80 41 47 0.0125 24 60 29 48 0.05 32 95 37 50 0.05 -7 95 0 51 0.05 17 97 23 52 0.0125 8 90 14 53 0.05 45 94 49 54 0.05 37 88 41 55 0.05 68 92 71 56 0.0125 28 75 33 57 0.05 61 92 64 58 0.025 46 64 50 59 0.05 1 93 8 60 0.0125 33 50 38 61 0.05 61 96 63 62 0.05 60 95 63 63 0.05 37 94 42 CuCl 2 , 10 tM 7.0 ± 4.6 [0066] Example 12. Comparative activity of isolated metal-hydrazone complexes and parent hydrazones towards Chlamydomonas reinhardtii and Synechocystis Hydrazones and their isolated metal complexes were compared with respect to their in vitro activity toward C. reinhardtii and the cyanobacterium (blue-green alga) Synechocystis sp. strain 6803. Metal complexes of hydrazones were prepared by precipitation from ethanol 37 WO 2012/024494 PCT/US2011/048260 with various metal salts at 1:1, 2:1 or 3:1 molar ratios, as described in general by Ainscough, Brodie, Dobbs, Ranford, and Waters (Inorganica Chimica Acta 1998, 267, 27-38, which is expressly incorporated by reference herein). [0067] A general synthesis of 1:1 metal-hydrazone complexes is as follows. The starting salicylaldehyde benzoylhydrazone or 2-hydroxyphenylketone benzoylhydrazone is dissolved (or suspended) in EtOH (generally 0.1 mmol hydrazone per mL solvent) and agitated at a temperature ranging from room temperature to 80 'C for 30 min. To this solution (or suspension) is added 1 equivalent of the metal salt (generally as a 1 M solution in EtOH). The mixture is agitated for a period ranging from 1 to 24 h at a temperature ranging from room temperature to 80 'C. The metal-hydrazone complex generally precipitates during the reaction or upon cooling and is isolated by filtration, washed with EtOH and finally washed with Et 2 0. In the instances where the complex does not precipitate, the solvent is removed and the resulting solid metal-hydrazone complex is washed with Et 2 O. Properties of particular metal complexes of hydrazones have been described (PCT Patent Application PCT/US10/021040 filed on 14 January 2010). Table 5. Analytical Data for Complexes Parent Number Comaond Metal Salt Ratio* Appearance mp (C) Number 72 1 Cu(OCOCH 3

)

2

-H

2 0 1:1 dark green 73 1 Cu(OCOCH 3

)

2

-H

2 0 2:1 tan solid 310-312 74 1 CuSO4-5H20 2:1 dark green 310-312 1 CuS 4 *5H 2 2:1solid 75 1 CuCl2-2H20 1:1 light green 311-312 olive solid 76 1 CuCl 2 -2H 2 0 2:1 light green 288-290 solid mustard 77 1 MnCl 2 -4H 2 0 2:1 colored 250 solid 38 WO 2012/024494 PCT/US2011/048260 Parent oper Comaond Metal Salt Ratio* Appearance mp ('C) Number 78 1 ZnCl 2 2:1 yellow- 250 green solid 25 79 7 CuC2-2H20 1:1 green 298-299 6 CuCl 2 -2H 2 0 1:1 solid 80 6 CuCl 2 -2H 2 0 1:1 olive green 215-219 solid 81 63 CUC1 2 *2H 2 0 1:1 olive green 273-274 solid 82 63 FeCl 3 -6H 2 0 1:1 black olid 199-202 83 63 FeCl 3 -6H 2 0 2:1 dark green 258-260 solid 84 63 FeCl 3 -6H 2 0 3:1 dark brown 264-268 solid 85 12 CuCl 2 -2H 2 0 1:1 olive green 242-247 solid 86 14 CuCl 2 -2H 2 0 1:1 olive green 283-294 solid 87 15 CuCl 2 -2H 2 0 1:1 olive green 224-235 solid 88 14 CuCl 2 -2H 2 0 1:1 olive green 287-289 solid 89 21 CuCl 2 -2H 2 0 1:1 olive green 299-308 solid 90 25 CuCl 2 -2H 2 0 1:1 olive green 242-246 solid 39 WO 2012/024494 PCT/US2011/048260 Parent oper Cyaond Metal Salt Ratio* Appearance mp ('C) Number 91 27 CuC12-2H 2 0 1:1 olive green 280-286 solid 92 32 CuC12-2H 2 0 1:1 olive green 279-281 solid 93 45 CuC12-2H20 1:1 olive green 257-267 45 C~l 2 2H 2 01:1solid 94 52 CuC12-2H20 1:1 olive green 212-270 solid 95 56 CuC12-2H20 1:1 olive green 270-274 56 C~l 2 2H 2 01:1solid 96 64 CuC12-2H 2 0 1:1 olive green 226-228 solid 97 6 U12H011olive green 245-248 65 C~l 2 2H 2 01:1solid 98 66 CuC12-2H 2 0 1:1 olive green 279-281 solid 99 68 CuC12-2H 2 0 1:1 green solid 206-289 100 26 CuCl 2 -2H 2 0 1:1 olive green 236-239 solid *Molar ratio of hydrazone:metal used to prepare complexes. [0068] In vitro assays against C. reinhardtii were conducted as described in Example 10 using TAP Cu minus medium. Growth inhibition was determined by comparing growth in the presence of test compound with growth in control wells lacking test compound. [0069] Results for growth inhibition of C. reinhardtii by hydrazones and corresponding isolated metal complexes (each at 1 tg/mL) are shown in Table 6. The results 40 WO 2012/024494 PCT/US2011/048260 illustrate that isolated copper complexes of hydrazones have much higher activity against C. reinhardtii than the corresponding hydrazones and also are much more active than isolated Fe, Mn and Zn complexes of hydrazones. Table 6. Growth Inhibition of C. reinhardtii by hydrazones and isolated metal-hydrazone complexes Parent Hydrazone Compound Complex Hydrazone Complex Number Number Metal Salt Ratio* % Inhn. % Inhn. 1 72 Cu(OCOCH 3

)

2

-H

2 0 1:1 16 96 1 73 Cu(OCOCH 3

)

2

-H

2 0 2:1 16 96 1 74 CuSO 4 -5H 2 0 2:1 16 96 1 75 CuCl 2 -2H 2 0 1:1 16 95 1 76 CuCl 2 -2H 2 0 2:1 16 75 1 77 MnCl 2 -4H 2 0 2:1 16 -1 1 78 ZnCl 2 2:1 16 -5 7 79 CuCl 2 -2H 2 0 1:1 29 74 6 80 CuCl 2 -2H 2 0 1:1 13 72 63 81 CuCl 2 -2H 2 0 1:1 -18 68 63 82 FeCl 3 -6H 2 0 1:1 -18 23 63 83 FeCl 3 -6H 2 0 2:1 -18 0 63 84 FeCl 3 -6H 2 0 3:1 -18 -4 12 85 CuCl 2 -2H 2 0 1:1 4 52 14 86 CuCl 2 -2H 2 0 1:1 6 74 15 87 CuCl 2 -2H 2 0 1:1 11 73 18 88 CuCl 2 -2H 2 0 1:1 13 47 21 89 CuCl 2 -2H 2 0 1:1 6 71 25 90 CuCl 2 -2H 2 0 1:1 33 49 27 91 CuCl 2 -2H 2 0 1:1 30 94 32 92 CuCl 2 -2H 2 0 1:1 21 95 45 93 CuCl 2 -2H 2 0 1:1 9 95 52 94 CuCl 2 -2H 2 0 1:1 -4 96 56 95 CuCl 2 -2H 2 0 1:1 11 95 64 96 CuCl 2 -2H 2 0 1:1 -5 95 65 97 CuCl 2 -2H 2 0 1:1 -5 95 66 98 CuCl 2 -2H 2 0 1:1 -4 95 *Molar ratio of hydrazone:metal used to prepare complexes. 41 WO 2012/024494 PCT/US2011/048260 [0070] In vitro assays against Synechocystis were conducted as described in Example 11 using BG-11 Cu minus medium. Growth inhibition was determined by comparing growth in the presence of test compound with growth in control wells lacking test compound. [0071] Results for growth inhibition of Synechocystis by hydrazones and corresponding isolated metal complexes are shown in Table 7. The results illustrate that isolated copper complexes of hydrazones have much higher activity against Synechocystis than the corresponding hydrazones and also are much more active than isolated Fe, Mn and Zn complexes of hydrazones. Table 7. Growth Inhibition of Synechocystis by hydrazones and isolated metal-hydrazone complexes Parent Hydrazone Test Compound Complex Concn. Hydrazone Complex Number Number Metal Salt Ratio* pg/ml % Inhn. % Inhn. 1 72 Cu(OCOCH 3

)

2

-H

2 0 1:1 0.05 35 98 1 73 Cu(OCOCH 3

)

2

-H

2 0 2:1 0.05 35 97 1 74 CuSO 4 -5H 2 0 2:1 0.05 35 95 1 75 CuCl 2 -2H 2 0 1:1 0.05 35 97 1 76 CuCl 2 -2H 2 0 2:1 0.05 35 96 1 77 MnCl 2 -4H 2 0 2:1 0.05 35 35 1 78 ZnCl 2 2:1 0.05 35 59 7 79 CuCl 2 -2H 2 0 1:1 0.0125 62 85 6 80 CuCl 2 -2H 2 0 1:1 0.025 70 91 63 81 CuCl 2 -2H 2 0 1:1 0.05 23 96 63 82 FeCl 3 -6H 2 0 1:1 0.05 23 24 63 83 FeCl 3 -6H 2 0 2:1 0.05 23 45 63 84 FeCl 3 -6H 2 0 3:1 0.05 23 33 27 91 CuCl 2 -2H 2 0 1:1 0.05 41 95 32 92 CuCl 2 -2H 2 0 1:1 0.05 68 95 52 94 CuCl 2 -2H 2 0 1:1 0.05 45 96 68 99 CuCl 2 -2H 2 0 1:1 0.05 16 94 26 100 CuCl 2 -2H 2 0 1:1 0.05 50 91 *Molar ratio of hydrazone:metal used to prepare complexes. 42 WO 2012/024494 PCT/US2011/048260 [0072] Example 13. Effect of copper on activity of metal-hydrazone complexes towards Chlamydomonas reinhardtii The effect of copper on activity of metal-hydrazone complexes towards C. reinhardtii was evaluated by comparing their activity in TAP Cu minus and TAP Cu plus media as described in Example 10 (compound concentrations were selected which gave low inhibition in TAP Cu minus medium). Growth inhibition was determined by comparing growth in the presence of test compound with growth in control wells lacking test compound. [0073] Results for growth inhibition by test compounds in TAP Cu plus medium (% Inhn. Plus Copper Observed) were compared with predicted values (% Inhn. Plus Copper Predicted) that were calculated using the formula set forth by S. R. Colby in Weeds 1967, 15, 20-22 based on results obtained for the same compounds in TAP Cu minus medium (% Inhn. Minus Copper Observed) and the inhibition attributed to copper chloride alone, as determined by comparing growth in TAP Cu minus and TAP Cu plus media without any test compound across experiments. Data are presented in Table 8. Results show that algicidal activity of metal complexes of hydrazones towards C. reinhardtii is synergistically enhanced in the presence of added copper. Furthermore, the fungitoxicity of copper complexes of hydrazones is synergistically enhanced in the presence of added copper. Table 8. Growth Inhibition of C. reinhardtii %Inhn. % Inhn. Parent Minus Plus % Inhn. Hydrazone Comple Test Copper Copper Plus Compound x Conen. Observe Observe Copper Number Number Metal Salt Ratio* pg/mL d d Predicted 1 72 Cu(OCOCH 3

)

2

-H

2 0 1:1 0.25 79 83 66 1 73 Cu(OCOCH 3

)

2

-H

2 0 2:1 0.25 23 87 10 1 74 CuSO 4 -5H 2 0 2:1 0.25 -9 82 -22 1 76 CuCl 2 -2H 2 0 2:1 0.5 3 89 -10 1 75 CuCl 2 -2H 2 0 1:1 0.25 18 59 5 63 82 FeCl 3 -6H 2 0 1:1 0.5 -16 89 -29 63 83 FeCl 3 -6H 2 0 2:1 0.5 -2 58 -15 63 84 FeCl 3 -6H 2 0 3:1 0.5 -19 70 -32 1 77 MnCl 2 -4H 2 0 2:1 0.5 -5 93 -18 1 78 ZnCl 2 2:1 0.5 -9 87 -22 CuCl 2 , -12.9 10 PM *Molar ratio of hydrazone:metal used to prepare complexes. 43

Claims (12)

1. A synergistic mixture for controlling the growth of algae, the synergistic mixture including copper and a hydrazone compound of Formula 1: X 3 Y 3 X4 OH O24 ) N, N W Y5 X 5 H n X 6 R Y 6 Formula 1 wherein W is -CHR 1-; n is 0 or 1; R is H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 3 -C 6 cycloalkyl, C 1 -C 6 haloalkyl, C 2 -C 6 haloalkenyl, C 2 -C 6 haloalkynyl, or C 3 -C 6 halocycloalkyl; RI is H, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 3 -C 6 cycloalkyl, C 1 -C 6 haloalkyl, C 2 -C 6 haloalkenyl C 2 -C 6 haloalkynyl, C 3 -C 6 halocycloalkyl, substituted aryl, unsubstituted aryl, substituted heteroaryl, or unsubstituted heteroaryl; X 3 , X 4 , X 5 , and X 6 are each independently selected from the group consisting of H, halogen, nitro, hydroxyl, cyano, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 C 4 alkylthio, C 1 -C 4 haloalkyl, C 1 -C 4 haloalkoxy, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, C 1 -C 4 haloalkylthio, -SO 2 R1, -SONRIR1, -CR1=NOR1, -CONRIR1, -NR1COOR1, -COOR1, substituted aryl, substituted heteroaryl, unsubstituted aryl, and unsubstituted heteroaryl; and Y 2 , Y 3 , Y 4 , Y 5 , and Y 6 are each independently selected from the group consisting of H, halogen, nitro, hydroxyl, cyano, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, C 2 -C 4 alkenyl, C 2 -C 4 alkynyl, C 1 C 4 alkylthio, C 1 -C 4 haloalkyl, C 1 -C 4 haloalkoxy, C 2 -C 4 haloalkenyl, C 2 -C 4 haloalkynyl, C 1 -C 4 haloalkylthio, -SO 2 R1, -SONRIR1, -R1=NOR1, -CONRIR1, -NR1COOR1, -COOR1, NR1R1, substituted aryl, substituted heteroaryl, unsubstituted aryl, unsubstituted heteroaryl, and phenoxy; 44 WO 2012/024494 PCT/US2011/048260 with the proviso that X 3 and X 4 , X 4 and X 5 , X 5 and X 6 , Y 2 and Y 3 , or Y 3 and Y 4 may form a 5- or 6-membered fused ring which may contain up to two heteroatoms selected from the group consisting of 0, N, and S.

2. Use of the synergistic mixture of claim 1 for controlling the growth of algae.

3. Use of the synergistic mixture of claim 1 for controlling the growth of algae in freshwater or marine environments, or on inert substrates selected from the group consisting essentially of wood, metal, and plastic.

4. Use of the synergistic mixture of claim 1 for controlling the growth of algae belonging to at least one of green algae and blue-green algae (cyanobacteria).

5. The synergistic mixture of claim 1 wherein the algae is selected from the group consisting of Chlamydomonas reinhardtii, Chlorella pyrenoidosa, Scenedesmus quadricauda, Chlorococcum oleofaciens, Selenastrum species, Phormidium species, Anabaena flosaquae, Nostoc commune, Osiffiatorae species, Synechocystis species, Synechococcus species; and Dunaliella parva..

6. The synergistic mixture of claim 1, wherein a growth inhibiting amount of the hydrazone compound of Formula I in mixture with copper is provided as a mixture in which the total molar ratio of copper to the hydrazone compound of Formula 1 exceeds 1:1.

7. The synergistic mixture of claim 1, wherein a growth inhibiting amount of the hydrazone compound of Formula I is provided as an isolated hydrazone-copper complex in which the molar ratio of the copper to the hydrazone compound of Formula 1 is one of 1:1 and 1:2.

8. The synergistic mixture of claim 1, wherein the hydrazone compound of Formula 1 to be combined with copper is complexed with a metal.

9. The synergistic mixture of claim 8, wherein the metal complexed with the hydrazone 2+ 2+ compound of Formula 1 is selected from the group consisting essentially of Cu', Cu , Fe Fe3 , Zn2+, and Mn2+ 45 WO 2012/024494 PCT/US2011/048260

10. The synergistic mixture of claim 1, wherein the copper is provided as at least one of the group consisting of copper oxychloride, copper octanoate, copper ammonium carbonate, copper arsenate, copper oxysulfate, copper formate, copper propionate, copper oxyacetate, copper citrate, copper chloride, copper diammonium chloride, copper nitrate, copper carbonate, copper phosphate, copper pyrophosphate, copper disodium EDTA, copper diammonium EDTA, copper oxalate, copper tartrate, copper gluconate, copper glycinate, copper glutamate, copper aspartate, copper adipate, copper palmitate, copper stearate, copper caprylate, copper decanoate, copper undecylenate, copper neodecanoate, copper linoleate, copper oleate, copper borate, copper methanesulfonate, copper sulfamate, copper acetate, copper hydroxide, copper oxide, copper oxychloride-sulfate, copper sulfate, basic copper sulfate, copper-oxine, copper 3-phenylsalicylate, copper chloride hydroxide, copper dimethyldithiocarbamate, ammonium copper sulfate, copper magnesium sulfate, copper naphthenate, copper ethanolamine, chromated copper arsenate, ammoniacal copper arsenate, ammoniacal copper zinc arsenate, ammoniacal copper borate, Bordeaux mixture, copper zinc chromate, cufraneb, cupric hydrazinium sulfate, cuprobam, nano-copper materials, copper didecyldimethylammonium chloride, Algimycin P11-C, Aquatrine, A&V-70, Cutrine-plus, Stocktrine II and K-Tea algicide and where appropriate the hydrates of such compounds.

11. The synergistic mixture of claim 1, wherein a ratio of the hydrazone to the copper is from 1:0.1 to 1:10,000.

12. An algicidal composition including the synergistic mixture of claim 1 and at least one of a fungicide, a herbicide, an insecticide, a bacteriocide, a nematocide, a miticide, a biocide, a termiticide, a rodenticide, a molluscide, an arthropodicide, a fertilizer, a growth regulator, and a pheromone. 46